Each CRISPR/Cas9 Knockout (KO) Plasmid product consists of a pool of 3 plasmids designed to ensure identification and cleavage of a specific gene for maximum knockout efficiency. Each plasmid encodes a unique 20 nt sequence derived from the GeCKO (v2) library.
Nek4 CRISPR Plasmids (h)
- Target species: human; for corresponding mouse product, see Nek4 CRISPR Plasmids (m)
- CRISPR/Cas9 KO Plasmids consists of Nek4-specific 20 nt guide RNA sequences derived from the GeCKO (v2) library
- For CRISPR gene knockout, gRNA sequences direct the Cas9 protein to induce a site-specific double strand break (DSB) in the genomic DNA
- Target-specific CRISPR Plasmids for both gene knockout and activation are available. Please refer to the detailed product information in the tabs below
- Gene knockdown or activation can be assayed using Nek4 Antibody (2631C1a): sc-81332
- All products are provided as transfection-ready, purified plasmid DNA, except the Lentiviral Activation Particles which have been prepackaged as Lentiviral Particles for use with hard-to-transfect cells
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Ordering Information
CRISPR/Cas9 KO Plasmid
HDR Plasmid
Double Nickase Plasmids
CRISPR Activation Plasmids
Lentiviral Activation Particles
Recommended CRISPR and HDR Support Reagents
Recommended Control Products
SEE ALSO...
Nek4 Antibodies for analysis of cellular responses to Nek4 CRISPR Products
Learn more about the CRISPR System
CRISPR/Cas9 KO plasmids, Double Nickase Plasmids and CRISPR/dCas9 Activation plasmids are considered a "Licensed Product" and are to be used in accordance with the Limited/Label License. Click here for details
Description
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nek4 CRISPR/Cas9 Knockout (KO) Plasmid (h2) consists of a pool of three plasmids each encoding the Cas9 nuclease and a Nek4-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency
- gRNA sequences are derived from the GeCKO (v2) library and direct the Cas9 protein to induce a site-specific double strand break (DSB) in the genomic DNA
- For selection of cells containing a successful Cas9-induced DSB, co-transfection with Nek4 HDR Plasmid (h2) (sc-405220-HDR-2) is recommended
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Nek4 Antibody (2631C1a): sc-81332
See Protocol Online.
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Description
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nek4 HDR Plasmid (h2) is recommended for co-transfection with Nek4 CRISPR/Cas9 KO Plasmid (h2)
- Nek4 HDR Plasmid (h2) consists of a pool of 2-3 plasmids, each containing a homology-directed DNA repair (HDR) template corresponding to the cut sites generated by the Nek4 CRISPR/Cas9 KO Plasmid (h2)
- Each HDR plasmid contains two 800 bp homology arms designed to specifically bind to the genomic DNA surrounding the corresponding Cas9-induced double strand DNA break site
- Each HDR Plasmid inserts a puromycin resistance gene to enable selection of stable knockout (KO) cells and an RFP (Red Fluorescent Protein) gene to visually verify transfection
- The puromycin resistance and RFP genes are flanked by two LoxP sites to allow for further processing by the Cre Vector sc-418923
See Protocol Online.
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Description
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nek4 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- The paired gRNAs encoded by Nek4 Double Nickase Plasmid (h2) differ from the paired gRNAs in Nek4 Double Nickase Plasmid (h)
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Nek4 Antibody (2631C1a): sc-81332
See Protocol Online.
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Description
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Nek4 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Nek4 CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- Following transfection, gene activation efficiency can be assayed by WB, IF or IHC using antibody: Nek4 Antibody (2631C1a): sc-81332
See Protocol Online.
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Description
- 200 µl of transduction-ready, high-titer CRISPR/dCas9 Lentiviral Activation Particles
- Nek4 Lentiviral Activation Particles (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically and efficiently upregulate gene expression via lentiviral transduction of cells
- Nek4 Lentiviral Activation Particles (h) contain the following SAM Activation elements: a deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, an MS2-p65-HSF1 fusion protein and a target-specific 20 nt. guide RNA. They also contain the blasticidin, hygromycin and puromycin resistance genes
- Upon transduction, the SAM complex binds to a site-specific region approximately 200-250 nt. upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- The guide RNA in the Nek4 Lentiviral Activation Particles (h2) encodes a target-specific 20 nt guide RNA that differs from the guide RNA in Nek4 Lentiviral Activation Particles (h)
- Following transfection, gene activation efficiency can be assayed by WB, IF or IHC using antibody: Nek4 Antibody (2631C1a): sc-81332
See Protocol Online.
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Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Nek4 CRISPR/Cas9 KO Plasmid (h2) | sc-405220-KO-2 | 20 µg | $390.00 | |||
Nek4 HDR Plasmid (h2) | sc-405220-HDR-2 | 20 µg | $437.00 | |||
Nek4 Double Nickase Plasmid (h) | sc-405220-NIC | 20 µg | $402.00 | |||
Nek4 Double Nickase Plasmid (h2) | sc-405220-NIC-2 | 20 µg | $402.00 | |||
Nek4 CRISPR Activation Plasmid (h) | sc-405220-ACT | 20 µg | $390.00 | |||
Nek4 CRISPR Activation Plasmid (h2) | sc-405220-ACT-2 | 20 µg | $390.00 | |||
Nek4 Lentiviral Activation Particles (h) | sc-405220-LAC | 200 µl | $447.00 | |||
Nek4 Lentiviral Activation Particles (h2) | sc-405220-LAC-2 | 200 µl | $447.00 |